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  Delivery of mtZFNs into Early Mouse Embryos

McCann, B. J., Cox, A., Gammage, P. A., Stewart, J. B., Zernicka-Goetz, M., & Minczuk, M. (2018). Delivery of mtZFNs into Early Mouse Embryos. Methods Mol Biol, 1867, 215-228. doi:10.1007/978-1-4939-8799-3_16.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0004-7213-6 Version Permalink: http://hdl.handle.net/21.11116/0000-0004-73A8-D
Genre: Journal Article

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McCann, B. J.1, Author
Cox, A.1, Author
Gammage, P. A.1, Author
Stewart, J. B.1, Author
Zernicka-Goetz, M.1, Author
Minczuk, M.1, Author
Affiliations:
1Stewart – Mitochondrial Mutations and Genome Co-evolution, Research Groups, Max Planck Institute for Biology of Ageing, Max Planck Society, Joseph-Stelzmann-Str. 9b, D-50931 Cologne, DE, ou_1942301              

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Free keywords: Germline In vitro transcription Micromanipulation Mitochondrial disease mtZFN
 Abstract: Mitochondrial diseases often result from mutations in the mitochondrial genome (mtDNA). In most cases, mutant mtDNA coexists with wild-type mtDNA, resulting in heteroplasmy. One potential future approach to treat heteroplasmic mtDNA diseases is the specific elimination of pathogenic mtDNA mutations, lowering the level of mutant mtDNA below pathogenic thresholds. Mitochondrially targeted zinc-finger nucleases (mtZFNs) have been demonstrated to specifically target and introduce double-strand breaks in mutant mtDNA, facilitating substantial shifts in heteroplasmy. One application of mtZFN technology, in the context of heteroplasmic mtDNA disease, is delivery into the heteroplasmic oocyte or early embryo to eliminate mutant mtDNA, preventing transmission of mitochondrial diseases through the germline. Here we describe a protocol for efficient production of mtZFN mRNA in vitro, and delivery of these into 0.5 dpc mouse embryos to elicit shifts of mtDNA heteroplasmy.

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 Dates: 20182018
 Publication Status: Published in print
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 Identifiers: Other: 30155826
DOI: 10.1007/978-1-4939-8799-3_16
ISSN: 1940-6029 (Electronic)1064-3745 (Linking)
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Title: Methods Mol Biol
Source Genre: Journal
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Pages: - Volume / Issue: 1867 Sequence Number: - Start / End Page: 215 - 228 Identifier: -